EP0202269B1 - Analysiervorrichtung zum optischen nachweis von gelösten stoffen - Google Patents

Analysiervorrichtung zum optischen nachweis von gelösten stoffen Download PDF

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Publication number
EP0202269B1
EP0202269B1 EP85905623A EP85905623A EP0202269B1 EP 0202269 B1 EP0202269 B1 EP 0202269B1 EP 85905623 A EP85905623 A EP 85905623A EP 85905623 A EP85905623 A EP 85905623A EP 0202269 B1 EP0202269 B1 EP 0202269B1
Authority
EP
European Patent Office
Prior art keywords
probe
fiber
signal
test signal
analytical instrument
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired
Application number
EP85905623A
Other languages
English (en)
French (fr)
Other versions
EP0202269A1 (de
Inventor
Anthony Ringrose
Ranald M. Sutherland
Claus Dähne
John F. Place
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Prutec Ltd
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Prutec Ltd
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Filing date
Publication date
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Publication of EP0202269A1 publication Critical patent/EP0202269A1/de
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/648—Specially adapted constructive features of fluorimeters using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/7703—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator using reagent-clad optical fibres or optical waveguides
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N2021/6484—Optical fibres
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N2021/7769—Measurement method of reaction-produced change in sensor
    • G01N2021/7786—Fluorescence

Definitions

  • the present invention concerns an analytical apparatus for optically determining species in solution, more especially for determining bioactive species by reactions of the immunoassaytype.
  • Analytical apparatuses comprising optical fiber probes which can optically monitor the adsorption of chemical species on the fiber core are known.
  • This technique is based on the immersion of a lit optical wave guide, for instance an optical fiber without cladding, in a test solution the refractive index of which is lower than that of the fiber core, whereby an interaction takes place between the evanescent wave component of the beam travelling along the guide and some species in solution to be determined.
  • This approach is particularly interesting for monitoring events in the reaction space in close vicinity to the fiber, i.e.
  • the light probe (optical fiber the cladding of which has been at least partly removed) is connected (coupled) by its proximal end to an optical excitation-detection arrangement comporting generally a light source providing an excitation signal of wavelength ⁇ 1' a beam splitter mirror separator to direct the signal into the proximal end of the fiber and a photodetector oriented at an angle with the source for receiving the test signal returned from the fiber, the beam splitter mirror effecting the separation between incident and returned light signals.
  • the return signal is a fluorescent signal generated by some component of the species under reaction with a wavelength ⁇ 2 longer than ⁇ 1 . Therefore the beam splitter mirror is preferably a dichroic mirror, i.e. a low pass interference filter with cut-off frequency between the maximum absorption and maximum fluorescence back emission of the fluorophore of interest. This arrangement allows a rather clean separation between the two signals.
  • this is generally constituted by a piece of optical fiber deprived from its normal low refractive index cladding.
  • This probe is generally coated with a film of one of the reactive species involved in an immunotype reaction, e.g. an antibody specific to an antigen to be analyzed.
  • an immunotype reaction e.g. an antibody specific to an antigen to be analyzed.
  • the antigen reacts on the fiber surface to provide an immunotype complex capable of generating a fluorescence signal when excited by the evanescent wave component, this signal being returned to the detector via the fiber and the optical coupling system.
  • a perturbing signal may however be produced from the interaction of the excitation signal exiting from the fiber end and the bulk of the analyte and to avoid this drawback the fiber tip has been either blackened or made fully reflective.
  • both the incident signal and the forward generated signal are absorbed with consequent decreased response and in the second case both the incident and fluorescent signals are returned to the separator with consequent relative high noise to signal ratio.
  • the apparatus represented in Figure 1 comprises essentially a probe 1 to be immersed in a container 2 containing an analyte solution.
  • the probe 1 is a piece of optical fiber of which the cladding has been removed on nearly its full extent; a small portion designated 1a still has the cladding not removed for mechanical coupling convenience as seen later.
  • the lower unclad portion of the fiber dipping in the sample liquid is coated with a layer of a reactive material susceptible to bind with the analyte dissolved in the sample to be analyzed in container 2, thus providing a complex on the surface of the guide which will fluoresce at wavelength ⁇ 2 (test signal).
  • the present apparatus further comprises an optical elements arrangement comprising a light source 3, focussing means 4, a beam separating means 5, a test signal detector 6, a reference signal detector 7 and a section of flexible optical fiber 8 held by positioning means 9 in proper optical position relative to the focussing means for receiving the excitation signal at wavelength ⁇ 1 from source 3 and at a proper angle for ensuring propagation by multiple reflection along the fiber 8.
  • an optical elements arrangement comprising a light source 3, focussing means 4, a beam separating means 5, a test signal detector 6, a reference signal detector 7 and a section of flexible optical fiber 8 held by positioning means 9 in proper optical position relative to the focussing means for receiving the excitation signal at wavelength ⁇ 1 from source 3 and at a proper angle for ensuring propagation by multiple reflection along the fiber 8.
  • detectors 6 and 7 are normally suitably connected to circuitry for processing the detected signals, amplifying, discriminating and calculating results in the form of electric signals to be displayed as readouts on suitable display means, all such technology being well known from people skilled in the art.
  • the flexible fiber element 8 is coupled to the probe by coupling means 10 shown in more details on Figure 2.
  • Such means comprise essentially three balls 11 (only two are shown) housed in a frusto-conically shaped recess 12 of a coupler
  • Probe 1 is therefore easily detachable by pulling off the coupler and interchangeably replaceable by a fresh one when used up after one analytical operation.
  • a dichroic mirror is provided atthe distal end of fiber 1 as shown in Figure 3.
  • This dichroic mirror comprises three layers of transparent materials 21, 22, 23 having suitable refractive indexes to filter out the incident wavelength ⁇ 1 and freely pass the test signal wavelength ⁇ 2 .
  • the manner in which to select the material with suitable refractive indexes is well known (se for instance Applied optics and optical Engineering, Editor R. Kingslake, Vol. 1, pages 316-322, Academic Press, New-York).
  • the mirror further comprises a metallic fully reflective back layer 24 (for instance of polished aluminum) which will reflect the ⁇ 2 signal backwards into the probe and from there back to the detector 6.
  • the distal end of the probe comports an optical filter of a substance capable of selectively absorbing the signal at wavelength ⁇ 1 and freely passing the test signal at wavelength ⁇ 2.
  • the arrangement is completed, like in the previous embodiment, by a polished fully reflective metallic mirror 26.
  • Narrow band filters of the type used in this embodiment are well known and available commercially from optical components manufacturers.
  • the probe 1 bare section is first coated with a reactive component susceptible to selectively bind some kind of analyte to be tested.
  • fluorescent labelling is provided in connection with either one or the other of the reactants, being provided however that no fluorescence (or only reduced fluorescence) is produced in the absence of the desired immunoreaction product.
  • the coated probe after suitable rinsing and drying, is connected to the flexible section 8 by means of the coupler 10.
  • the optical components 3 to 7 and the additional electronics are turned on and the lit probe is immersed into the container 2 containing the analyte to be determined by the specific aforementioned reaction on the probe surface.
  • a fluorescence test signal of wavelength X 2 is generated at the probe surface by the interaction of the excitation light evanescent component of wavelength ⁇ 1 and the analyte complex layer.
  • the backward component of the testsignal is directly returned along the fiber toward the detector 6 via fiber 8, and splitter 5 whereas the forward component first reaches the distal end of the fiber where it is back reflected by the mirror 24, 26 after traversing the fitering section (see Figures 3 and 4) inserted between the fiber tip and the mirror.
  • the unwanted excitation component travelling forward is eliminated by absorption. This technique enables to improve the signal to noise ratio at the detector 6 and enables to increase sensitivity with a given light intensity input or to reduce the input energy without sacrificing sensitivity.
  • the probe of the present instrument can be directly introduced, in situ, into the bloodstream of the patient or, if not possible, into a by-pass circuit temporarily connected, for instance during surgery.
  • the probe of the apparatus of the invention is constituted by optical and mechanical elements of sufficiently small size to be used directly in such tests with only a minimum of discomfort for the . patient.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Plasma & Fusion (AREA)
  • Engineering & Computer Science (AREA)
  • Optics & Photonics (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Optical Measuring Cells (AREA)

Claims (5)

1. Analysevorrichtung für immunologische Untersuchungen, bestehend aus einer optischen Fasersonde, die an ein optisches System gekoppelt ist, das aus einer Erregungssignalquelle, einem Testsignaldetektor und einem strahlenzerlegenden Bauteil zum Trennen des Testsignals einer Wellenlänge Ä2 vom dem Erregungssignal der Wellenlänge À1 besteht, wobei die Faser mit einem bioreaktiven Reaktionsmittel beschichtet ist, das befähigt ist, sich spezifisch mit einem Analyten zu verbinden, der in einer Probenlösung gelöst ist, um einen für immunologische Untersuchungen typischen Komplex zu bilden, wobei durch Wechselwirkung zwischen dem Erregungssignal, das der Sonde aufgedrückt wird, und diesem Komplex ein fluoreszierendes Testsignal erzeugt wird, wobei das Testsignal über die verspiegelte Spitze der Sonde, die Kupplung und das Trennbauteil zum Sammeln reflektierend auf den Detektor zurückgeworfen wird, wodurch es die gewünschte analytische Information liefert, dadurch gekennzeichnet, daß die Fasersondenspitze mit Mitteln zum Blockieren des Einfallssignals und zum selektiven Hindurchlassen und Zurücksenden des Testsignals zu dem Detektor versehen ist.
2. Analysevorrichtung nach Anspruch 1, wobei die Mittel zum Reflektieren des Testsignals und zum Blockieren von λ1 und zum Hindurchlassen von À2 aus einem an der Spitze der Sonde angebrachten dichroitischen Filterspiegel bestehen.
3. Analysevorrichtung nach Anspruch 1, wobei die Mittel zum Blockieren von λ1 und zum Hindurchlassen von Ä2 aus einem optischen Absorptionsfilter bestehen, das zwischen dem Faserende und der verspiegelten Sodenspitze befindlich ist.
4. Analysevorrichtung nach Anspruch 1, weiterhin bestehend aus einem flexiblen optischen Faserabschnitt zwischen der Sonde und dem System, wobei die Sonde mit diesem Abschnitt mittels einer steckerförmigen einrastenden Kupplung lösbar verbunden ist.
5. Analysevorrichtung nach Anspruch 4, wobei die einrastende Verbindung aus einem hohlen Stecker besteht, der ständig auf ein Ende des flexiblen Faserabschnittes aufgesetzt ist, wobei der Stecker am entgegengesetzten Ende eine Öffnung zum Einsetzen des freien Endes der Sonde aufweist und wobei in einer Ausnehmung des Steckers eine in Kommunikation mit der Öffnung gehaltene, aus einer Feder und Kugeln bestehende Klemmvorrichtung vorgesehen ist, die auf das nahegelegene.Ende der Sonde einwirkt, um sie in optisch gekoppelter Position mit dem flexiblen Abschnitt zu halten.
EP85905623A 1984-11-06 1985-11-04 Analysiervorrichtung zum optischen nachweis von gelösten stoffen Expired EP0202269B1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CH5306/84 1984-11-06
CH5306/84A CH660633A5 (fr) 1984-11-06 1984-11-06 Appareil d'analyse pour la determination optique de substances en solution.

Publications (2)

Publication Number Publication Date
EP0202269A1 EP0202269A1 (de) 1986-11-26
EP0202269B1 true EP0202269B1 (de) 1989-01-25

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Application Number Title Priority Date Filing Date
EP85905623A Expired EP0202269B1 (de) 1984-11-06 1985-11-04 Analysiervorrichtung zum optischen nachweis von gelösten stoffen

Country Status (7)

Country Link
EP (1) EP0202269B1 (de)
JP (1) JPH065213B2 (de)
AU (1) AU578331B2 (de)
CH (1) CH660633A5 (de)
DE (1) DE3567968D1 (de)
ES (1) ES8705629A1 (de)
WO (1) WO1986003004A1 (de)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3953163B1 (de) 2019-04-11 2023-10-11 Compagnie Générale des Etablissements Michelin Vollreifen aus elastomerem werkstoff für eine seilbahnrolle

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4852967A (en) * 1986-03-25 1989-08-01 Ciba Corning Diagnostics Corp. Evanescent wave sensors
JPH01267443A (ja) * 1988-04-19 1989-10-25 Mitsubishi Kasei Corp 光学的測定方法及び装置
US4925268A (en) * 1988-07-25 1990-05-15 Abbott Laboratories Fiber-optic physiological probes
US5401469A (en) * 1989-04-19 1995-03-28 Ibiden Co., Ltd. Plastic optical biomaterials assay device
US5173747A (en) * 1990-09-20 1992-12-22 Battelle Memorial Institute Integrated optical directional-coupling refractometer apparatus
US5377008A (en) * 1990-09-20 1994-12-27 Battelle Memorial Institute Integrated optical compensating refractometer apparatus
JP3107649B2 (ja) * 1991-12-20 2000-11-13 イビデン株式会社 蛍光免疫測定装置
US5354574A (en) * 1992-06-23 1994-10-11 Ibiden Co., Ltd. Method for producing optical fiber having formyl groups on core surface thereof
EP1126278B1 (de) * 1992-07-31 2007-05-23 Inverness Medical - BioStar Inc. Vorrichtungen und Verfahren zur Detektion eines Analyten mittels optischer Interferenz
JPH07110330A (ja) * 1993-09-01 1995-04-25 Bio Sensor Kenkyusho:Kk 光学センサー
JP2706616B2 (ja) * 1994-02-04 1998-01-28 株式会社バイオセンサー研究所 液体の光学的測定装置
FR2846420B1 (fr) * 2002-10-24 2006-05-05 Commissariat Energie Atomique Dispositif de lecture de luminescence integre

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3947088A (en) * 1974-08-22 1976-03-30 Weber Dental Manufacturing Co. Interface for light probe
US4356396A (en) * 1980-12-17 1982-10-26 Siemens Corporation Fiber optical measuring device with compensating properties
US4582809A (en) * 1982-06-14 1986-04-15 Myron J. Block Apparatus including optical fiber for fluorescence immunoassay

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3953163B1 (de) 2019-04-11 2023-10-11 Compagnie Générale des Etablissements Michelin Vollreifen aus elastomerem werkstoff für eine seilbahnrolle

Also Published As

Publication number Publication date
ES548538A0 (es) 1987-05-01
EP0202269A1 (de) 1986-11-26
AU578331B2 (en) 1988-10-20
AU5067085A (en) 1986-06-03
JPH065213B2 (ja) 1994-01-19
ES8705629A1 (es) 1987-05-01
WO1986003004A1 (en) 1986-05-22
CH660633A5 (fr) 1987-05-15
DE3567968D1 (en) 1989-03-02
JPS62501102A (ja) 1987-04-30

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